发状念珠藻NfwspA3基因的克隆及其耐旱能力分析北大核心CSCD

水分胁迫蛋白(WSP,Water stress protein,)在植物耐干旱过程中起着重要作用。发状念珠藻具有极强的耐旱、耐盐和抗辐射等能力。本研究以发状念珠藻基因组DNA为模板,利用PCR技术克隆到一个发状念珠藻水分胁迫蛋白基因的开放阅读框(ORF)序列,命名为NfwspA3,其序列全长1017 bp。生物信息学预测表明,该基因可编码338个氨基酸残基,预测等电点为4.85,分子量为37.48 kD,为亲水性稳定蛋白,分布在细胞质中。构建了NfwspA3基因的重组表达菌株pET32aNfwspA3,以聚乙二醇-6000(PEG-6000)对其模拟干旱胁迫,发现重组菌株最高可忍耐60%浓度的PEG-6000,表现出较强的耐旱性,且具有较高的半乳糖苷酶活性。研究结果为探知发状念珠藻WSP蛋白逆境响应机制及其遗传改良提供了理论依据和基因元件。     

不同温度对念珠藻生理生化特性的影响

文章尝试比较不同温度对念珠藻（Nostocsp.）细胞生长和生理特性的影响。主要内容是分别在10℃、25℃、35℃和40℃环境中对念珠藻进行9 d的室内培养，然后测定不同温度下念珠藻的生物量、光合活性、总多糖、总蛋白质和总脂质含量。研究结果表明，低温（10℃）对念珠藻细胞生长的促进与光合活性的提高基本无影响；而高温（35℃、40℃）会促进藻细胞的生长和光合活性的提升。也就是说，低温时念珠藻细胞的多糖含量几乎不变，蛋白质含量与脂质含量先升高后降低；而高温时多糖含量和蛋白质含量增加，脂质含量降低。以上结果说明温度胁迫会使念珠藻对生物大分子物质进行调控用于抵抗环境变化。     

发状念珠藻干旱胁迫响应基因NfGR的克隆与鉴定

该研究采用PCR技术,从发状念珠藻细胞中克隆了谷胱甘肽还原酶(glutathione reductase,GR)基因,命名为NfGR,其开放阅读框长1 374bp,编码458个氨基酸,蛋白相对分子量为49.42kD,理论等电点为5.49。氨基酸序列分析表明,NfGR蛋白具有NADPH结合位点超家族(NADB-Rossmann superfamily)和吡啶氧化还原酶二聚体超家族(Pyr_redox_dim superfamily)2个结构域,与点形念珠藻(Nostoc punctiforme)的相似性达93%。系统进化树分析表明,NfGR与点形念珠藻处在同一进化枝上,亲缘关系较近。qRT-PCR表达分析表明,在不同浓度PEG-6000处理下,NfGR基因均保持上调表达,其中,PEG-6000浓度为8%时,NfGR基因的相对表达量达到峰值(32.69)。研究推测,谷胱甘肽还原酶可能参与了发状念珠藻对干旱胁迫过程的响应。     

发状念珠藻藻殖段的分化及其光合特性的研究

发状念珠藻 (NostocflagelliformeBorn .etFlah .)存在着两个重要而明显的个体发育阶段 ,即营养藻丝体和藻殖段。采用弱光 (铺垫砂粒遮光 ) ,红光或在白光下向培养基中加入DCMU (3,4_dichlorophenyl_1,1_dimethylurea)等方法 ,可促进营养藻丝体转变成藻殖段。用可见光吸收光谱、低温荧光发射光谱和光合放氧活性表示发状念珠藻藻丝体与藻殖段的光合特性 ,表明营养藻丝体和藻殖段的可见光吸收光谱和色素含量差别不大。而两者在不同光强范围 (110～ 12 0 0 μmol·m-2 ·s-1)和不同温度 (15～ 45℃ )下的光合放氧活性 ,表明发状念珠藻的藻殖段比营养藻丝体可能更适合在低光强下和较高的温度下生长。从荧光发射光谱可以看出 ,在光合能量传递中营养藻丝体比藻殖段在两个光系统之间的光能分配上更加均衡 ;但是藻殖段中藻胆体吸收光能向两个光系统的传递比营养藻丝体的更加有效。可以认为藻殖段的形成对光合作用的结构与功能产生影响。     

温度和初始pH对生物结皮中4种优势蓝藻生长的影响

生物土壤结皮对干旱半干旱地区生态环境健康及可持续发展有重要意义。具鞘微鞘藻、伪枝藻、念珠藻和鱼腥藻是生物结皮中蓝藻的优势种,对其大规模培养有着广阔的应用前景。以4种蓝藻为材料,采用室内液体培养方法,研究了5个温度(10、20、25、30、35℃)和5个溶液初始pH(4、6、8、10、12)对4种藻生物量和溶液pH的影响。结果表明：在20℃以下温度培养,供试4种藻生物量增长缓慢,在25～35℃下4种藻均能正常生长,其中25和30℃下藻类生物量增长最快。不同藻种的最适培养温度略有区别,伪枝藻和念珠藻最适培养温度为25～30℃,具鞘微鞘藻和鱼腥藻为30℃。供试4种藻对溶液pH有极强的调节能力,均能够在5个初始pH环境中进行增殖,在初始pH为4的培养环境下能达到最大生物量和比生长速率,在初始pH为12下生物量和比生长速率最低。研究为生物结皮中优势藻的扩繁提供了科学依据。     

利用16S rRNA基因分析3种念珠藻的亲缘关系

通过基因克隆和序列测定,获得了地木耳、发菜和葛仙米的16S rRNA序列。以集胞藻为外类群,采用邻接法(NJ)和最大简约法(MP),对14株念珠藻的16S rRNA基因序列进行系统发育分析,两种方法得到的系统发育树基本一致。结果显示:(1)三者间的遗传距离(0.0129~0.0261,平均值0.0203)均大于普通念珠藻的种内遗传距离(0.0007~0.0118,平均值0.0074),超过了种内差异程度,在系统树中这3种念珠藻分别聚簇在不同的分支上;(2)地木耳、发菜和葛仙米与裂褶念珠藻等其它7种念珠藻间的遗传距离(0.0423~0.0996,平均值0.0561)大于三者间的遗传距离,在系统树中聚类较近,而与其它念珠藻聚类较远。据此,可将地木耳、发菜和葛仙米归为念珠藻属3个不同的种,并且它们的亲缘关系较近。     

异形胞分化相关基因在点形念珠藻厚壁孢子中的转录表达

点形念珠藻(Nostoc punctiforme)ATCC29133是一种丝状固氮蓝藻,能分化产生异形胞、厚壁孢子、藻殖段等细胞类型。异形胞是一种提供微氧条件以进行固氮作用的特化细胞,厚壁孢子是在逆境下产生的能耐受干旱、冰冻等恶劣环境的细胞1。与异形胞相似,厚壁孢子的外被也具有多糖和糖脂成分2,3。厚壁孢子在某些蓝藻藻丝       

念珠藻生长特性研究

研究了念珠藻的生长周期、固氮特性、生长条件等生长特性.得出本实验念珠藻可以固氮;磷能有效制约其生长;需要长时间的光照等结果.     

西北常见野菜地木耳的经济价值及其开发利用

地木耳是一种常见的广泛分布于我国西北干旱、半干旱地区的念珠藻属植物。概述了其较好的食用、药用保健价值以及非常独特的生理生态特征,并进一步指出了如何更好地开发利用这种植物资源,从而达到可持续利用。     

念珠藻属植物hetR基因的适应性进化分析

以念珠藻属(Nostoc)及其近缘类群hetR基因的51条序列为研究对象,对hetR基因的编码蛋白进行生物信息学分析和系统发育分析,并使用分支模型、位点模型和分支-位点模型进行该基因位点的适应性进化研究。系统发育分析结果显示,51条hetR基因蛋白序列可分为4个大分支。适应性进化分析结果表明,在3种进化模型中,大多数分支及藻株都没有检测到统计学上具有显著性的正选择位点,说明检测的位点大多处于负选择压力下。但在普通念珠藻(Nostoc commune,CHAB2802)中检测到正选择位点(126T),提示念珠藻属植物hetR基因发生了适应性改变。     

Developing sporophytes transition from an inducible to a constitutive ecological strategy of desiccation tolerance in the moss Aloina ambigua: effects of desiccation on fitness

Background and Aims Two ecological strategies of desiccation tolerance exist in plants, constitutive and inducible. Because of difficulties in culturing sporophytes, very little is known about desiccation tolerance in this generation and how desiccation affects sexual fitness.Methods Cultured sporophytes and vegetative shoots from a single genotype of the moss Aloina ambigua raised in the laboratory were tested for their strategy of desiccation tolerance by desiccating the shoot–sporophyte complex and vegetative shoots at different intensities, and comparing outcomes with those of undried shoot–sporophyte complexes and vegetative shoots. By using a dehardened clonal line, the effects of field, age and genetic variance among plants were removed.Key Results The gametophyte and embryonic sporophyte were found to employ a predominantly inducible strategy of desiccation tolerance, while the post-embryonic sporophyte was found to employ a moderately constitutive strategy of desiccation tolerance. Further, desiccation reduced sporophyte fitness, as measured by sporophyte mass, seta length and capsule size. However, the effects of desiccation on sporophyte fitness were reduced if the stress occurred during embryonic development as opposed to postembryonic desiccation.Conclusions The effects of desiccation on dehardened sporophytes of a bryophyte are shown for the first time. The transition from one desiccation tolerance strategy to the other in a single structure or generation is shown for only the second time in plants and for the first time in bryophytes. Finding degrees of inducible strategies of desiccation tolerance in different life phases prompts the formulation of a continuum hypothesis of ecological desiccation tolerance in mosses, where desiccation tolerance is not an either/or phenomenon, but varies in degree along a gradient of ecological inducibility.     

Inheritance of seed desiccation sensitivity in a coffee interspecific cross: evidence for polygenic determinism

The genetic determinism of seed desiccation sensitivity was studied using a cross between two coffee species exhibiting a large difference for this trait, Coffea pseudozanguebariae (tolerant) and C. liberica (sensitive). Throughout the whole study, seed desiccation tolerance was quantified both in terms of water content and water activity. Whatever the parameter used, the level of seed desiccation tolerance in F1 hybrids corresponded to that of the mid-parent, thus indicating an additive inheritance of seed desiccation tolerance at the F1 level. A broad variation was observed among hybrids backcrossed to C. liberica (BCs) for seed desiccation tolerance, independent of the parameter used to quantify it. This variation was continuous and BCs showed transgression in the direction of the most desiccation sensitive parent, indicating (i) that desiccation tolerance is a polygenic trait in coffee species, and (ii) that C. pseudozanguebariae does not present the most favourable alleles for all the genes involved in seed desiccation tolerance. No significant difference was observed between the two reciprocal backcrosses, F1xC. liberica and C. libericaxF1, for the level of desiccation tolerance of their seeds, showing the absence of a maternal effect on this trait. There was no significant effect of the number of seeds harvested from each BC on the level of desiccation tolerance of its seeds. Moreover, there was no significant correlation within BCs between seed size, seed viability, and water content before desiccation and desiccation tolerance.     

Desiccation Tolerance and Global Change: Implications for Tropical Bryophytes in Lowland Forests

Global change puts an increasing pressure on tropical forests and their inherent diversity by the risk of longer droughts and drier microclimatic conditions within the forest. How organisms will respond is uncertain, especially for organisms highly depending on their microclimatic environment such as bryophytes. An adequate tolerance to desiccation is important to face these changes, however, little is known for tropical bryophytes. We investigated for the first time the desiccation tolerance of epiphytic bryophytes from contrasting microsites at the tropical lowland forest in French Guiana. Using chlorophyll‐fluorescence (F_v/F_m) as an indicator of recovery, we tested: (1) desiccation tolerance for short (3 d) and long (9 d) desiccation events; (2) different desiccation intensities; and (3) recovery by rehydration with water vapor. Species from the canopy were well adapted to desiccation events. Thirteen of 18 species maintained more than 75 percent of their photosynthetic capacity after recovery at the strongest desiccation treatment of 9 d at 43 percent relative humidity (RH). In contrast, species from the understory were sensitive and withstood desiccation only at humid conditions of 75 percent RH and higher. The photosystem of the studied bryophytes was reactivated efficiently in equilibration with water vapor only—a yet neglected phenomenon in bryology. A novel introduced desiccation tolerance index allows global comparison of desiccation tolerances and highlights the sensitivity of understory species. Our results suggest that decreasing humidity caused by climate change and forest degradation could be a concerning threat for understory species.     

与种子耐脱水性有关的基础物质研究进展

耐脱水的获得和维持与种子的类型有关,正常型种子耐脱水,而顽拗形种子对脱水高度敏感,正常型种子的脱水耐性随发良[过程而变化,种子成熟时胚的脱水耐性增强,其萌发时胚变为不耐脱水,当种子获得脱水耐性时,糖,LEA蛋白质和抗氧化防御系统等保护性物质积累,但脱水耐性是一种复杂的数量的特性,任何一种单一的机制都不能 充分地解释脱水耐性,各种保护性物质协同调节脱水耐性,本文综述了近几年来关于种子耐脱水性与保护性物质相关性的研究进展。     

与种子耐脱水性有关的基础物质研究进展

耐脱水的获得和维持与种子的类型有关,正常型种子耐脱水,而顽拗形种子对脱水高度敏感。正常型种子的脱水耐性随发育过程而变化,种子成熟时胚的脱水耐性增强,而萌发时胚变为不耐脱水。当种子获得脱水耐性时,糖、LEA蛋白质和抗氧化防御系统等保护性物质积累。但脱水耐性是一种复杂的数量的特性,任何一种单一的机制都不能充分地解释脱水耐性,各种保护性物质协同调节脱水耐性。本文综述了近几年来关于种子耐脱水性与保护性物质相关性的研究进展。     

Tolerance of thermophilic and hyperthermophilic microorganisms to desiccation

We examined short- and long-term desiccation tolerance of 31 strains of thermophilic and hyperthermophilic Archaea and thermophilic phylogenetically deep-branching Bacteria. Seventeen organisms showed a significant high ability to withstand desiccation. The desiccation tolerance turned out to be species-specific and was influenced by several parameters such as storage temperature, pH, substrate or presence of oxygen. All organisms showed a higher survival rate at low storage temperatures (−20°C or below) than at room temperature. Anaerobic and microaerophilic strains are influenced negatively in their survival by the presence of oxygen during desiccation and storage. The desiccation tolerance of Sulfolobales strains is co-influenced by the pH and the substrate of the pre-culture. The distribution of desiccation tolerance in the phylogenetic tree of life is not domain specific. Surprisingly, there are dramatic differences in desiccation tolerance among organisms from the same order and even from closely related strains of the same genus. Our results show that tolerance of vegetative cells to desiccation is a common phenomenon of thermophilic and hyperthermophilic microorganisms although they originated from quite different non-arid habitats like boiling acidic springs or black smoker chimneys.     

Changes in sugars during rice seed desiccation

The correlation between desiccation tolerance and soluble sugars was investigated in seeds of a number of rice cultivars belonging to the Asian rice Oryza sativa L. They were dried or ultradried to various low moisture content and then imbibed for germination testing. Few or no changes on germination percentage and vigor index were found in Indian rice seeds even after their moisture content fell to 3.5%, indicating that Indian rice exhibited a strong desiccation tolerance. On the contrary, Japonica rice seed germination percentage rapidly decreased, after their moisture content fell to 4.5%. The capacity for desiccation tolerance in Japonica (cv. Chunjiang 15) and Indian (cv. Zhongzu 1) developing seeds increased on 23–40 and 15–25 days after pollination, respectively. Though the level of monosaccharides declined, the content of sucrose has increased during desiccation. These results suggest that desiccation tolerance might be associated with the increase in seed viability and the changes in sugar level, and that raffinose could be capable of contributing to the desiccation tolerance to ultradrying. Published in Russian in Fiziologiya Rastenii, 2006, Vol. 53, No. 2, pp. 220–226. The text was submitted by the authors in English.     

Stress-associated factors increase after desiccation of germinated seeds of <Emphasis Type="Italic">Tabebuia impetiginosa</Emphasis> Mart.

Improved re-establishment of desiccation tolerance was studied in germinated seeds of Tabebuia impetiginosa Mart. by exposing to a polyethylene glycol solution prior to desiccation. The effects of different osmotic potentials and drying rates were studied. In addition, temporary temperature stress and exogenous abscisic acid were applied to evaluate their effect on desiccation tolerance of the protruded radicle. An osmotic potential of −1.7 MPa at 5°C followed by slow drying was most effective in the re-establishment of desiccation tolerance in protruded radicles with a length up to 3 mm. An osmotic potential of −1.4 or −2.0 MPa was less effective. Fast drying completely prevented the re-induction of desiccation tolerance. Cold shock or heat shock prior to osmotic treatment as well as abscisic acid added to the osmotic solution improved desiccation tolerance of protruded radicles. Surprisingly, survival of the germinated seed did not depend on re-establishment of desiccation tolerance in the protruded radicle. Even after the protruded radicle became necrotic and died, the production of adventitious roots from the hypocotyls allowed for survival and the development of high quality seedlings. Thus, T. impetiginosa appeared to be well adapted to the seasonally dry biome in which the species thrives via mechanisms that offer protection against desiccation in the young seedling stage.     

Geographic variation in adult and embryonic desiccation tolerance in a terrestrial-breeding frog

Intraspecific variation in the ability of individuals to tolerate environmental perturbations is often neglected when considering the impacts of climate change. Yet this information is potentially crucial for mitigating deleterious effects of climate change on threatened species. Here we assessed patterns of intraspecific variation in desiccation tolerance in the frog Pseudophryne guentheri, a terrestrial-breeding species experiencing a drying climate. Adult frogs were collected from six populations across a rainfall gradient and their dehydration and rehydration rates were assessed. We also compared desiccation tolerance of embryos and hatchlings originating from within-population parental crosses from four of the populations. Embryos were reared on soil at three soil–water potentials and their desiccation tolerance was assessed across a range of traits. We found significant and strong patterns of intraspecific variation in almost all traits, both in adults and first-generation offspring. Adult frogs exhibited clinal variation in their water balance responses, with populations from drier sites both dehydrating and rehydrating more slowly compared to frogs from more mesic sites. Similarly, desiccation tolerance of first-generation offspring was significantly greater in populations from xeric sites. Our findings suggest that populations within this species will respond differently to the regional reduction in rainfall predicted by climate change models.